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1. (WO1991007422) METHOD AND KIT FOR PURIFYING NUCLEIC ACIDS
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/1991/007422 International Application No.: PCT/US1990/004276
Publication Date: 30.05.1991 International Filing Date: 31.07.1990
Chapter 2 Demand Filed: 10.05.1991
IPC:
C12N 1/06 (2006.01) ,C12N 15/10 (2006.01) ,C12Q 1/68 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1
Micro-organisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving micro-organisms or compositions thereof; Processes of preparing or isolating a composition containing a micro-organism; Culture media therefor
06
Lysis of micro-organisms
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
10
Processes for the isolation, preparation or purification of DNA or RNA
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
68
involving nucleic acids
Applicants:
BOEHRINGER MANNHEIM CORPORATION [US/US]; 9115 Hague Road Indianapolis, IN 46250, US
Inventors:
REARDON, Melissa; US
KLEIN, Lisa; US
Agent:
HANSON, Norman, D.; Felfe & Lynch 805 Third Avenue New York, NY 10022, US
Priority Data:
434,32413.11.1989US
Title (EN) METHOD AND KIT FOR PURIFYING NUCLEIC ACIDS
(FR) PROCEDE ET KIT DE PURIFICATION D'ACIDES NUCLEIQUES
Abstract:
(EN) The invention teaches a method and kit for purifying nucleic acid, such as DNA, from a sample, such as lysed cell or tissue sample. A sample is appplied to an anionic exchange matrix column uniformly distributing the sample therein. The column bed is then washed with a weak ionic salt solution which is then removed. The anionic exchange material is optionally primed with an amount of strong ionic salt solution which is insufficient to elute the nucleic acid from the column. The priming solution is then removed. An elution buffer is added either directly after the washing step or after the priming step. This is also a strong ionic salt solution of an ionic salt. The elution buffer removes the purified nucleic acid from the material. The method permits purification of nucleic acids without using organic solvents, and, if the priming step is used, in more concentrated form. Uniform distribution of the sample via disturbance of the matrix or column facilitates the purification.
(FR) L'invention concerne un procédé et un kit de purification d'acides nucléiques, tels que l'ADN, à partir d'un échantillon, tel qu'un échantillon de tissu ou de cellules ayant subi une lyse. Un échantillon est appliqué à une colonne de matrice échangeuse d'anions distribuant uniformément l'échantillon à l'intérieur de la colonne. Le lit de la colonne est ensuite lavé avec une solution d'un sel ionique faible qui est ensuite éliminé. Le matériau échangeur d'anions est éventuellement amorcé avec une quantité d'une solution d'un sel ionique fort qui est insuffisante pour effectuer l'élution de l'acide nucléique de la colonne. La solution d'amorçage est ensuite éliminée. Un tampon d'élution est ajouté soit directement après l'étape de lavage soit après l'étape d'amorçage. Ce tampon est également une solution d'un sel ionique fort d'un seul sel ionique. Le tampon d'élution élimine l'acide nucléique purifié du matériau échangeur d'anions. Le procédé permet la purification d'acides nucléiques sans l'utilisation de solvants organiques et, si l'étape d'amorçage est appliquée, dans une forme plus concentrée. La distribution uniforme de l'échantillon par perturbation de la matrice ou colonne facilite la purification.
Designated States: JP
European Patent Office (AT, BE, CH, DE, DK, ES, FR, GB, IT, LU, NL, SE)
Publication Language: English (EN)
Filing Language: English (EN)
Also published as:
EP0508985